The rodipicolinate synthase gene was cloned from Nicotiana tabacum L.cv,which encodes the key enzyme in the synthesis of lysine,and then a vitro molecular reform reliefing feedback suppression was made. Some resistant seedlings were acquired using the mutated gene as selectable marker and analog of lysine as selectable reagent,and the detection results were positive by means of PCR and Real-Time PCR,however the phenotype is somewhat abnormal.

Objective To explore the clinical application and outlook of anterior segment observation with Stratus optical coherence tomography(Stratus OCT).Design Prospective case series.Participants 56 eyes in 38 patients with primary angle closure glaucoma (PACG).Methods Morphological changes of anterior chamber angle (ACA) were observed with Stratus OCT before and 4 weeks after laser iridectomy.Opening status of ACA were described and measured with Photoshop software.Main Outcome Measures Opening degree of ACA and ratio of anterior chamber depth to thickness of cornea.Results To a great deal of extent,Stratus OCT could be used to observe the appearance of ACA. Clear images of ACA could be acquired.The ACA before laser was (15.67?5.33) degree and significantly reopened to(26.56?8.17) degree after laser iridectomy (P=0.000).The ratio of anterior chamber depth to thickness of cornea was also changed from (0.39?0.13) pre-treatment to (0.89?0.32) pest-treatment(P=0.000).Conclusions Stratus OCT could be applied to observe the changes of opening degree of ACA easily.The standard of quantization should be consummated in the future.

Objective Understand the variation of serum creatinine measurement in clinical laboratories of some hospitals in Beijing.Methods 8 samples of mixed frozen human serum added different creatinine concentration standard materials(the creatinine concentration were80-1 000 ?mol/L)and 8 samples of mixed frozen serum of patients(contained different creatinine concentration)were distributed to 13 clinical laboratories(31 series of detection systems)with the way of spot investigation.Every clinical laboratories measured the samples followed the standard operating procedure.Results As to the mixed frozen human serum added different creatinine standard materials,the CV of different detection systems results were 5.74%-9.68%;as to the mixed frozen patients' serum,the CV was 5.90%-11.69%. Compared with Beckman closed detection systems,the results of Dade systems(which used the kinetic alkaline pieric acid method)showed the bias were-5.99%-0.35%,and as to the other systems which measured by alkaline picric acid method,when creatinine concentrations were 200 ?mol/L,the results showed negative bias,and the greatest bias was-8.45%.The bias plots revealed negative for all of the detection systems with enzymatic method over the whole concentration range,and the greatest bias was -8.88%.Conclusions The creatinine determination results of Beckman and Dade closed detection systems were consistent.The results of detection systems which used enzymatic method were generally lower than Beckman detection systems.What's more,the creatinine measurement variations of clinical laboratories were very large,especially for the results of unclosed detection systems,so it was urgent need to solve the standardization of creatinine measurement.

Animals ; Dose-Response Relationship, Drug ; Hydroxylamine ; toxicity ; Male ; Rats ; Rats, Wistar ; Spermatozoa ; drug effects ; Testis ; cytology ; drug effects

Objective To elucidate the roles of TANK-binding kinase-1(TBKl)in hepatitis B virus (HBV)infection induced interferon antiviral immunity.Methods Peripheral blood monocytes were separated by CD14 magnetic microbeads from healthy volunteers(HV)and chronic hepatitis B(CHB)patients.Purified mDCs were induced and proliferated in the culture medium with human granulocyte-macrophage concentration of 25 mg/L were stimulated.The mRNA expressions of TBK1,interferon regulatory factor (IRF)3 and interferon(IFN)-βwere quantified by real time polymerase chain reaction(PCR).The levels of IFN-β in supernatants were determined by enzyme-linked immunosorbent assay(ELISA).Reslllts The mRNA levels of TBK1,IRF3 and IFN-β did not change significantly at 0,12,24 and 48 h after the significantly at 0, 12, 24 and 48 h in CHB group, whereas, it was significantly up-regulated at 12 h in HV group. Conclusions Our results suggest that there may be some disorders in host antiviral signal transduction pathways downstream the binding between ligands and receptors on mDC surface. The insufficient IFN-β expression after HBV infection may result in persistent chronic infection.

Objective To investigate the expression and function of retinoic acid-inducible gene Ⅰ(RIG-Ⅰ) in monocyte-derived dendritic cells (MoDC) at different stages of hepatitis B virus(HBV)infection and to explore the role of RIG-Ⅰ in the disease progression after HBV infection. Methods Peripheral blood samples were collected from 28 hepatitis B virus-infected persons, including 21 cases of chronic hepatitis B (CHB) and 7 of acute hepatitis B (AHB). Eighteen healthy subjects were recruited as controls. Purified CD14~+ monocytes were isolated by CD14 microbeads. MoDCs were induced from CD14~+ monocytes with granulocyte macrophage-colony stimulating factor (GM-CSF) and interleukin (IL)-4 for 7 days, and then were infected with vesicular stomatitis virus (VSV) to stimulate RIG-Ⅰ expression. The mRNA expression levels of RIG-Ⅰ, interferon (IFN )-promoter stimulating factor-1 (IPS-1) and IFN-β at 16 hours and 24 hours after infection with VSV were measured by real-time quantitative polymerase chain reaction (PCR). Data with normal distribution were tested by analysis of variance. Continuous variables between groups were compared using Mann-Whitney U test. Comparison among multiple groups was done by Kruskal-Wallis test. Results The expression levels of RIG-Ⅰ in MoDCs from CHB patients were significantly lower than those in AHB patients and healthy controls at 16 hours (2.44±2.03, 19. 54±3. 15, 21. 48±8. 39, respectively; F=7.451,P=0.002) and 24 hours (2. 68±2. 93, 10. 31 ±3. 88, 14. 01 ±5. 04, respectively, F = 7. 908, P = 0. 001)following VSV stimulation. The IPS-1 levels in both CHB patients and AHB patients were higher than those in healthy controls at 16 hours (2. 05±l. 08, 1. 99±1. 56, 0. 60±0. 31, respectively) F=7.246,P =0.003) and 24 hours (2. 27±2. 16, 3.24 ± 1.21, 1. 08±0. 73, respectively; F= 13. 598, P = 0. 001).Furthermore, the IFN-β expression levels were significantly lower in CHB patients compared to AHB patients and healthy controls at 16 hours and 24 hours after VSV stimulation. Conclusions The expressions of RIG-Ⅰ and IPS-1 in MoDC are abnormal in HBV infected persons, which indicates that RIG-Ⅰ signaling pathway might be blocked by HBV. The impaired function of MoDC may play a role in HBV infection and chronicity.

AIM:To investigate the safety and effect of femtosecond laser-assisted cataract surgery with Cionni modified capsular tension ring (MCTR) implantation in the management of traumatic lens subluxation.METHODS: Totally 11 patients (11 eyes) with traumatic lens subluxation were divided into three groups according to the severity of lens dislocation, ranging from 90° to 120° (4 eyes), 120° to 180° (5 eyes) and 180° to 270° (2 eyes).The contact LenSx femtosecond laser cataract surgery platform was applied to create the capsulotomy, prepare nuclear fragmentation and make corneal wound creation.Anterior vitrectomy was performed in some patients during the surgery.After capsular retractors insertion and phacoemulsification, the MCTR was inserted to the capsular bag and fixed to the sclera.Finally, the IOL was implanted into the capsular bag.Postoperative visual acuity, intra-and post-operative complications, anterior capsular opening, IOL and MCTR position and intraocular pressure (IOP) were assessed.RESULTS:The duration of follow-up was 2mo.All the operations were completed successfully.Five eyes underwent cataract surgery combined with anterior vitrectomy.Four eyes had been inserted with 2-eyelet MCTR and seven eyes with 1-eyelet MCTR.The best corrected visual acuity (BCVA) after operation was better than 0.5 in 4 eyes, between 0.3 and 0.5 in 3 eyes, between 0.1 and 0.3 in 3 eyes, and less than 0.1 in 1 eye.Compared with preoperative BCVA, the difference was statistically significant (P<0.05).All the IOLs were stably centered and the eyelet of MCTR was fixated steadily between the iris and the anterior capsule.The common intra-and post-operative complications were subconjunctival hemorrhage, incomplete capsulotomy, residual cortex, secondary glaucoma and posterior capsular opacification.CONCLUSION:Femtosecond laser-assisted cataract surgery can improve the success rate of capsulorhexis, and reduce the difficulty of nuclear fragmentation.Femtosecond laser-assisted cataract surgery combined with MCTR implantation is an ideal surgical method for traumatic lens subluxation.

AIM: To compare the accuracy of intraocular lens(IOL)power calculations by using five formulas(Haigis, SRK-T, Hoffer Q, Holladay-1, SRK-Ⅱ)in eyes with long axial lengths in order to improve the accuracy of predicating IOL powers.METHODS: Fifty-one eyes of 51 cases of age-related cataract and with mild long axial(24.5mm27mm) were collected who`s optical biometry were performed by the Zeiss IOL Master500 before operation.They underwent regular phacoemulsification and posterior chamber IOL implantation.The actual postoperative refraction was measured with the methods of phoropter and subjective optometry 3mo after surgery.Then we compared the differences of the predicted and actual postoperative refraction of the five formulas in each group.RESULTS: In the mild axial lengths cases, the differences between SRK Ⅱ formula and the other four formulas were statistically significant (P<0.05), and the difference between Hoffer Q and SRK-T formula was statistically significant (P<0.05);there was no difference among the other formulas (P>0.05).In the moderate and severe long axial lengths cases, the differences between SRK Ⅱ formula and the other four formulas were statistically significant (P<0.05), and the difference between Hoffer Q and SRK-T formula, Hoffer Q and Haigis formula were statistically significant (P<0.05);there was no difference among the other formulas (P>0.05).The differences of all the five formulas between the two groups were statistically significant (P<0.05).CONCLUSION: In the mild axial lengths cases, Haigis, SRK-T, Hoffer Q, Holladay-1 performed well.In the moderate and severe long axial lengths cases, Haigis, SRK-T and Holladay-1 performed better than other formulas.The accuracy of all the five formulas decreases as the axial length getting longer.

Objective To obtain acquired immunity evidence in hamsters elicited by third stage hookworm larvae of Necator americanas(NaL3).morphology changes of NaL3 and inflammatory responses in the skin and undedying subcutaneous tissue and muscles of hamsters were observed.Methods Hamsters were immunized subcutaneously with one dose of 150 NaL3 at 2 weeks earlier,and then challenged pereutaneously with 900 NaL3.Skins were excised from post-challenge hamsters at 6,24,72 hours and 1,2 weeks,and then examined under light microscopy.Non-immunized hamsters served as negative controls.Results In non-immunized hamsters the number of NaL3 were 15,33,11.0 and 0 at 6,24,72 hours and 1,2 weeks post-infection.No damaged or dead NaL3 section was observed.All NaL3 exhibited no structural damage and infihrating inflammatory cells were absent from the sunDunding tissues.There were no cutaneous changes.In contrast.the total number of Nak sections in the skin of immunized hamsters were 25,53,15,5 and 4 at 6,24,72 hours and 1,2 weeks post-challenge.Among these NaL3 sections,damaged and dead section number were 0,24,6,0,0 and 0,0,7,5,4.At 24 hours post-challenge the Nak exhibited cutieular swelling and damage.By 72 hours post-challenge pyknosis of the somatic cells nuclei and sparseness or loss of definition in the internal structures of NaL3 were seen.One or two weeks after chanenge,the NaL3 showed severe damage or even dead with remnants.Inflammatory responses including macrophages,epithelioid cells and eosinophils infiltrating and granulomata forming were mainly seen around the NaL3 sections in the skin of immunized hamsters.Conclusions Hamsters initially immunized with NaL3 exhibited obvious acquired immunity protection against percutaneously challenged infection as evidenced by vigorous inflammatory responses in the skin and underlying subcutaneous tissue and muscle.

Objective To investigate the feature of diabetic foot ulcer and the relative mechanism, and to analyze its predilection site. Methods The precipitating factors and the areas of ulcer were analyzed in 121 diabetic patients with foot ulcer. The peripheral neuropathy was diagnosed by measuring the skin sensation, tendon reflex, electromyogram or vibration perception threshold (VPT). The diabetic peripheral neuropathy (DPN) was evaluated by positive symptoms, ultrasound or magnetic resonance angiography. According to morbid changes, the patients were divided into neuropathic ulcer group (n = 33), ischemic ulcer group (n = 27), and mixed type ulcer group (n = 61). Results The ulcers of 97 cases (80. 2%) were distributed in digital pedis and plantar pedis. In neurogenic ulcer group, there were 48.5% lesion at planta pedis, 30.3% at dorsum pedis, and 21.2% at digiti pedi. In ischemie ulcer group, there were 59.3% at digiti pedis, 29.6% at dorsum pedis, and 11.1% at planta pedis. In mixed type ulcer group, there were 49. 2% at digiti pedis, 41% at planta pedis, and 9.8% at dorsum pedis. The difference of ulcer distribution was apparent in these groups. Among these three groups, patients in mixed type ulcer group was the oldest, and had the longest duration of diabetes (P<0.05) and the largest area of ulcer at diagnosis. However, the BMI and blood pressure in isehemic ulcer group were higher than those of the other groups (P>0.05). Conclusions The difference of lesion basis affects the position of diabetic foot ulcer, and the thumbs and plantar pedis are the predilection area of diabetic foot.